欢迎访问过程工程学报, 今天是

›› 2008, Vol. 8 ›› Issue (4): 784-788.

• 生化工程专栏 • 上一篇    下一篇

红平红球菌LSSE8-1中辅因子再生系统的构建

李海波 邢建民 熊小超 刘会洲   

  1. 中国科学院过程工程研究所 中国科学院过程工程研究所分离科学与工程青年实验室 中国科学院过程工程研究所分离科学与工程实验室 中国科学院过程工程研究所分离工程与工程青年实验室
  • 收稿日期:2008-04-09 修回日期:2008-05-14 出版日期:2008-08-20 发布日期:2008-08-20
  • 通讯作者: 李海波

Construction of an Efficient Cofactor-regeneration System in Rhodococcus erythropolis LSSE8-1

LI Hai-bo XING Jian-min XIONG Xiao-chao LIU Hui-zhou   

  1. Institute of process engineering, Chinese Academy of Sciences Lab. Sep. Sci. & Eng., Institute of Process Engineering, Chinese Academy of Sciences Laboratory of Separation Science and Engineering, Institute of Process Engineering) Chinese Academy of Sciences Institute of Process Engineering, Chinese Academy of Sciences
  • Received:2008-04-09 Revised:2008-05-14 Online:2008-08-20 Published:2008-08-20
  • Contact: LI Hai-bo

PDF

被引次数

5

摘要： 采用PCR方法从红球菌Rhodococcus sp. RHA1基因组中克隆了编码甲酸脱氢酶的基因(fdh)，将该基因片段插入大肠杆菌-红球菌穿梭质粒pBS306，构建了甲酸脱氢酶表达质粒pBS-PFG，转入专一性脱硫菌R. erythropolis LSSE8-1，得到重组菌LSSE8-1-FDH. 2,3,5-三苯基氯化四氮唑(TTC)显色反应测得重组菌LSSE8-1-FDH的总脱氢酶活(OD496)为0.464，原始菌LSSE8-1的总脱氢酶活(OD496)为0.353，重组菌比原始菌酶活增加了31%. 考察了不同浓度甲酸根对R. erythropolis LSSE8-1与重组菌LSSE8-1-FDH生长的影响，当甲酸根浓度为25 mmol/L时重组菌LSSE8-1-FDH的生长最佳. 油水相静息细胞脱硫实验表明，重组LSSE8-1-FDH菌比宿主菌的脱硫速率增加了12.5%，总脱硫率增加了约20%.

关键词: 甲酸脱氢酶, 微生物脱硫, 红球菌, NADH, 二苯并噻吩

Abstract: The aim of this study was to develop a new biocatalyst with high NADH regeneration, which was helpful to increase desulfurizatin activity. Formate dehydrogenase gene (fdh) was amplified from Rhodococcus RHA1 and then introduced into an specific desulfurization bacterium, Rhodococcus erythropolis LSSE8-1, using an E. coli-R. erythropolis shuttle vector pBS306. TTC reaction showed that the dehydrogenase activity of LSSE8-1-FDH (OD496) was 0.464, the dehydrogenase activity of LSSE8-1 (OD496) was 0.353, and the dehydrogenase activity increased 31%. Recombinant strain showed a moderately faster growth rate in 25 mmol/L sodium formate solution. Desulfurization of DBT in the oil-aqueous two phases system by LSSE8-1 and LSSE8-1-FDH was conducted respectively. Compared with the orginal strain LSSE8-1, the recombinant strain LSSE8-1-FDH had an increase of 12.5% in desulfurizing activity, 4 h in desulfurizing time, and total desulfuriztion capacity increased 20%.

Key words: formate dehydrogenase, biodesulfuriztion, Rhodococcus erythropolis, NADH, dibenzothiophene

中图分类号: